title: Sample 6
description: Protocols: CSF1-cultured macrophages were collected and sorted into CD11b+CD206high populations which then were separated by each genotype based on the expression of CD45.1 or CD45.2. Bone marrow cells from CD45.1 and Clec4a2-/- (CD45.2) mice were co-cultured with 100 ng/ml CSF1 for 10 days. Total RNA was isolated by using the RNeasy micro kit (Qiagen). Total RNA quantity and integrity were assessed using Quant-IT RiboGreen RNA Assay Kit (ThermoFisher Scientific) and high-sensitivity DNA tapestation (Agilent). Purification of mRNA, generation of double stranded cDNA and library construction were performed using NEBNext Poly(A) mRNA Magnetic Isolation Module (New England Biolabs) and NEBNext Ultra II Directional RNA Library Prep Kit (New England Biolabs) for Illumina with adapters and barcode tags (dual indexing). The final size distribution of the pool was determined using high-sensitivity DNA tapestation (Agilent), and quantification was determined by Quant-IT RiboGreen RNA Assay Kit (ThermoFisher Scientific).
attributes.tag: ENA first public
attributes.value: 2021-10-28
attributes.tag: ENA last update
attributes.value: 2021-10-28
attributes.tag: External Id
attributes.value: SAMEA9418377
attributes.tag: INSDC center alias
attributes.value: Kennedy Institute of Rheumatology University of Oxford
attributes.tag: INSDC center name
attributes.value: Kennedy Institute of Rheumatology University of Oxford
attributes.tag: INSDC first public
attributes.value: 2021-10-28T05:45:20Z
attributes.tag: INSDC last update
attributes.value: 2021-10-28T05:45:20Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-10734:Sample 6
attributes.tag: age
attributes.value: 12
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: bone marrow macrophage
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: genotype
attributes.value: Clec4a2 -/-
attributes.tag: immunophenotype
attributes.value: CD11b+CD206high
attributes.tag: organism part
attributes.value: bone marrow
attributes.tag: sample name
attributes.value: E-MTAB-10734:Sample 6
attributes.tag: sex
attributes.value: male
attributes.tag: strain
attributes.value: C57BL/6J